Advanced Microscopy Techniques

In addition to the microscopes listed here, there are a number of advanced microscopy systems that have been developed by the MIN Director that can be used on a trail basis. These instruments are custom designed and built by Dr. Field and graduate students from Prof. Randy Bartels’ lab in the Electrical and Computer Engineering Department. Since these instruments are not part of the MIN, the preferred method for use is to contact both Dr. Field and Prof. Bartels to arrange a time to discuss your interests and potential collaborative efforts between your lab and the Bartels lab

Multiphoton Laser Scanning Microscopy

A custom multiphoton laser scanning microscope system designed and constructed by Dr. Field is located in the Scott Bioengineering building in Prof. Bartels’ laboratory. The unique construction of this microscope enables a larger field of view than is available with conventional laser-scanning microscopy platforms. Available contrast methods include two-photon excited fluorescence and second- and third-harmonic generation. There is no incubation stage on this microscope, and the upright geometry makes this system difficult to use with samples immersed in a liquid medium. The preferred sample preparation method is fixed tissues sliced to 250 µm or thinner and mounted on a standard 1″ x 3″ microscope slide. Details of this microscope, including additional photos, operating principles, and representative data can be found in an invited tutorial article: Michael D. Young, Jeffrey J. Field, Kraig E. Sheetz, Randy A. Bartels, and Jeff Squier, “A pragmatic guide to multiphoton microscope design,” Adv. Opt. Photon.  7, 276-378 (2015). An example image of two-photon excited fluorescence from a coronal slice of fixed murine cortical tissue (provided by Prof. Shane Hentges) is shown below.

coronal slice of fixed murine cortical tissue
Two-photon excited fluorescence from RFP bound to RNA (green) and unidentified endogenous fluorophores (purple). This image is a composite formed from numerous images to form a mosaic. The full size of the image is approximately 18000 x 12000 pixels, and the spatial resolution across the image is less than 1 µm.